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Elife. 2019 May 24;8. pii: e44635. doi: 10.7554/eLife.44635.

Comprehensive substrate specificity profiling of the human Nek kinome reveals unexpected signaling outputs.

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Department of Biology, Massachusetts Institute of Technology, Cambridge, United States.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States.
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, United States.
MIT Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, United States.
Department of Pharmacology, Yale School of Medicine, New Haven, United States.
Department of Medical Biophysics, University of Toronto, Toronto, Canada.
Biotech Research and Innovation Center, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Princess Margaret Cancer Center, University Health Network, Toronto, Canada.
Department of Surgery, Beth Israel Deaconess Medical Center, Divisions of Acute Care Surgery, Trauma, and Critical Care and Surgical Oncology, Harvard Medical School, Boston, United States.


Human NimA-related kinases (Neks) have multiple mitotic and non-mitotic functions, but few substrates are known. We systematically determined the phosphorylation-site motifs for the entire Nek kinase family, except for Nek11. While all Nek kinases strongly select for hydrophobic residues in the -3 position, the family separates into four distinct groups based on specificity for a serine versus threonine phospho-acceptor, and preference for basic or acidic residues in other positions. Unlike Nek1-Nek9, Nek10 is a dual-specificity kinase that efficiently phosphorylates itself and peptide substrates on serine and tyrosine, and its activity is enhanced by tyrosine auto-phosphorylation. Nek10 dual-specificity depends on residues in the HRD+2 and APE-4 positions that are uncommon in either serine/threonine or tyrosine kinases. Finally, we show that the phosphorylation-site motifs for the mitotic kinases Nek6, Nek7 and Nek9 are essentially identical to that of their upstream activator Plk1, suggesting that Nek6/7/9 function as phospho-motif amplifiers of Plk1 signaling.


Nek kinase family; Nima-related kinase; biochemistry; cancer biology; chemical biology; kinase biology; mitosis; none; signaling; substrate specificity

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